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| United States Patent |
5,124,352
|
|
Mizushima
, et al.
|
June 23, 1992
|
Fat emulsions containing isocarbacyclin
Abstract
The fat emulsions containing the prostaglandin I.sub.2 's expressed by the
following formula (I):
##STR1##
where X represents an oxygen atom to a methine group, Y is a carbon atom,
Z represents a methylene or methine group; when X is an oxygen atom, the
mode of Y-Z binding is a double bond of carbon-carbon, and when X is a
methine group, the mode of X-Y binding is a double bond of carbon-carbon
and Z is a methylene group; R.sub.1 represents a hydrogen atom or alkyl
group, R.sub.2 represents a hydrogen atom or fluorine atom, and R.sub.3
represents a hydrogen atom, methyl group, ethyl group or vinyl group.
R.sub.4 represents a substituted or unsubstituted alkyl group with 1-10
carbon atoms, a substituted or unsubstituted alkenyl group with 2-10
carbons atoms, a substituted or unsubstituted alkynyl group with 2-10
carbon atoms, or a substituted or unsubstituted cycloalkyl group with 3-8
carbon atoms; and n is zero (0) or an integer of value 1.
| Inventors:
|
Mizushima; Yutaka (25-20, Daia 4-chome, Setagaya-ku, Tokyo, 155, JP);
Shoji; Yoko (Kanagawa, JP);
Yagyu; Yasuko (Kanagawa, JP);
Kurozumi; Seizi (Tokyo, JP)
|
| Assignee:
|
Teijin Limited (Osaka, JP);
Mizushima; Yutaka (Tokyo, JP)
|
| Appl. No.:
|
727376 |
| Filed:
|
July 5, 1991 |
Foreign Application Priority Data
| Jun 17, 1985[JP] | 60-129920 |
| Current U.S. Class: |
514/510; 514/469; 514/573 |
| Intern'l Class: |
A61K 031/235 |
| Field of Search: |
514/469,510,573
|
References Cited
U.S. Patent Documents
| 4472428 | Sep., 1984 | Toru et al. | 549/214.
|
| 4613614 | Sep., 1986 | Fukaya et al. | 514/469.
|
| 4683633 | Aug., 1987 | Imagawa et al. | 514/573.
|
| Foreign Patent Documents |
| 0097481 | Jan., 1984 | EP.
| |
| 0132027 | Jan., 1985 | EP.
| |
| 53-50141 | May., 1978 | JP.
| |
| 59-210044 | Nov., 1984 | JP.
| |
| 210044 | Nov., 1984 | JP.
| |
| 60-13779 | Jan., 1985 | JP.
| |
| 60-149524 | Aug., 1985 | JP.
| |
| 60-169430 | Sep., 1985 | JP.
| |
| 60-243079 | Dec., 1985 | JP.
| |
| 60-260524 | Dec., 1985 | JP.
| |
| 61-44819 | Mar., 1986 | JP.
| |
| 2012168 | Aug., 1979 | GB.
| |
Other References
Hackh's Chemical Dictionary, McGraw-Hill 1972, p. 424.
|
Primary Examiner: Lee; Mary C.
Assistant Examiner: Haley; Jacqueline
Attorney, Agent or Firm: Sughrue, Mion, Zinn, Macpeak & Seas
Parent Case Text
This is a continuation of application Ser. No. 07/428,020, filed Oct. 26,
1989 abandoned which is a continuation of application Ser. No. 07/015,451
filed as PCT/JP86/00293, Jun. 12, 1986 now abandoned.
Claims
What is claimed is:
1. A pharmaceutical prostaglandin emulsion comprising:
a mixture of vegetable oil, phospholipid, water, and a pharmaceutically
effective amount of at least one compound of formula (Ia)
##STR5##
wherein R.sup.1 is selected from the group consisting of a hydrogen atom
and an unsubstituted alkyl group having 1 to 10 carbon atoms;
R.sup.21 is a hydrogen atom;
R.sup.3 is a hydrogen atom;
R.sup.4 is an unsubstituted alkyl group having 1-10 carbon atoms; and
n represents 0.
2. The pharmaceutical prostaglandin emulsion according to claim 1, wherein
R.sup.1 is a methyl group and R.sup.4 is a pentyl group.
3. The pharmaceutical prostaglandin emulsion according to claim 1, wherein
said mixture comprises 5-50% (w/v) vegetable oil, 1-50 parts of
phospholipid per 100 parts vegetable oil, from 1.0 mg/ml to 0.2 .mu.g/ml
of said compound of formula (Ia) and the balance water.
Description
TECHNICAL FIELD
This invention relates to fat emulsions containing the prostaglandin
I.sub.2 's.
More particularly, it relates to new fat emulsions containing, as an active
ingredient, the prostaglandin I.sub.2 's which are not only thrombolytic
but are useful in the treatment of cardiovascular-renal system disorders
as well.
BACKGROUND OF THE INVENTION
The prostaglandins exibit a wide range of physiological activities and have
been finding widespread medicinal applications because of their diverse
and useful biological actions such as peripheral circulatory improvement,
vasodilation, antiulceration, hypotensive, induction of labor,
thrombolytic, and antiasthmatic. In recent years, these compounds have
been studied for possible new indications such as anticancer,
osteometabolism improvement, antiviral, hepatic protection, diuresis In
particular, the naturally occurring prostacyclin is a local hormone
predominantly produced in vivo from the vascular wall of arteries; owing
to its potent physiological effects such as platelet agglutination
inhibitory activity and vasodilating activity, this local hormone is an
important factor which regulates in vivo cellular functions, and hence an
attempt has been made to use the naturally occurring prostacyclin per se
as a pharmaceutical product [P. J. Lewis, J. O. Grady et al., Clinical
prostacyclin, Raven Press, N.Y. (1981)].
On the other hand, however, when these useful prostaglandins are applied as
pharmaceutical products, various problems are encountered with respect to
the in vivo instability inherent in the prostaglandins, side effects
attributable to a wide range of their physiological effects, and the
difficulties of formulations due to their chemical instability.
Thus, intensive studies have been carried out at home and abroad with
regard to chemically stable synthetic prostacyclin derivatives comparable
to naturally occurring prostacyclin in terms of biological actions.
Meanwhile, attempts have been made to stabilize chemically unstable
prostacyclin in dosage form as well as to improve its drug efficacy. For
example, propositions have been put forth regarding a method of
stabilizing the prostacyclin as the clathrate compound using cyclodextrin
(Joseph Scesitri et al., Japan Laid-Open Patent Showa 54-56685), a method
of stabilizing the prostacyclin with surface active agents (Moo Yang Chou
et al., Japan Laid-Open Patent Showa 55-15470), a pharmaceutical
preparation by first obtaining a new ester derivative of prostacyclin with
the higher fat solubility, emulsifying this ester derivative in a fat, and
maintaining its activity comparable to that of the parent prostacyclin
(Fukaya et al., Japan Laid-Open Patent Showa 60-13779), and so forth.
The fat emulsions containing PGE.sub.1 or PGA.sub.1 have recently been
proposed as the stabilized prostaglandin fat preparations which possess
vasodilating, platelet agglutination inhibitory, and hypotensive
activities [Mizushima et al., Japan Laid-Open Patent Showa 58-222014 and
Japan Laid-Open Patent Showa 59-141518; and Mizushima et al., Ann. Rheum.
Diseases, 41, 263 (1982); Pharm. Pharmacol., 35, 398 (1983)]. Such
techniques are applied to the preparation of the anti-tumor agents; a
proposal has been set forth with respect to the improvement of selective
delivery of anticancer drugs to the target organ (Okamoto et al., Japan
Laid-Open Patent Showa 59-122423).
DISCLOSURE OF THE INVENTION
The inventors noticed the aforementioned facts and made intensive studies
on some of the chemically stable synthetic prostaglandin I.sub.2 's in
order to prolong their effects and enhance their clinical efficacy. As a
consequence, we prepared said fat emulsion containing the stable
prostaglandin I.sub.2 's, and discovered that the said preparations have
attained these objects. The inventors, therefore, arrived at the present
invention
This invention, therefore, concerns the fat emulsions containing the
prostaglandin I.sub.2 's expressed by the following formula (I):
##STR2##
where X represents an oxygen atom or a methine group, Y is a carbon atom,
Z represents a methylene or methine group; when X is an oxygen atom, the
mode of Y-Z binding is a double bond of carbon-carbon; and when X is a
methine group, the mode of X-Y binding is a double bond of carbon-carbon
and Z is a methylene group; R.sub.1 represents a hydrogen atom or alkyl
group, R.sub.2 represents a hydrogen atom or fluorine atom, and R.sub.3
represents a hydrogen atom, methyl group, ethyl group or vinyl group.
R.sub.4 represents a substituted or unsubstituted alkyl group with 1-10
carbon atoms, a substituted or unsubstituted alkenyl group with 2-10
carbons atoms, a substituted or unsubstituted alkynyl group with 2-10
carbon atoms, or a substituted or unsubstituted cycloalkyl group with 3-8
carbon atoms; and n is zero (0) or an integer of value 1.
The fat emulsions in the invention are new fat emulsions containing
chemically stable prostaglandin I.sub.2 's; such fat emulsions possess
longer duration of action, improve the stability of the prostaglandin
I.sub.2 's, reduce manifestation of side effects, exhibit, at the same
time, potent pharmacological effects, and are useful as the preparations
for use in intravenous administration.
Most of the prostaglandin I.sub.2 's in the aforementioned formula (I) are
the compounds known to be stabilized prostaglandin I.sub.2 's [Japan
Laid-Open Patent Showa 58-150583, Japanese Patent Application Number Showa
57-32981).]
In the aforementioned formula (I), X represents an oxygen atom or a methine
group, Y is a carbon atom, Z represents a methylene or methine group; when
X is an oxygen atom, the mode of Y-Z binding is a double bond of
carbon-carbon, and when X is a methine group, the mode of X-Y binding is a
double bond of carbon-carbon and Z is a methylene group. From these
definitions, it is preferable in the present invention that the active
ingredient is the isocarbacyclins expressed by the forllowing formula
(Ia):
##STR3##
(where R.sub.1, R.sub.3, R.sub.4, and n are defined as above; R.sub.21
represents a hydrogen atom); or that the active ingredient is the
7-fluoroprostacyclins expressed by the following formula (Ib):
##STR4##
(where R.sub.1, R.sub.3, R.sub.4, and n are defined as above; R.sub.22
represents a fluorine atom).
In the aforementioned formula (1), R.sub.1 is a hydrogen atom or alkyl
group. The said alkyl group may be an alkyl group with 1-10 carbon atoms
such as methyl, ethyl, propyl, butyl, pentyl, hexyl, octyl, and decyl
group. The R.sub.1 is preferably a hydrogen atom or methyl group.
R.sub.4 represents a substituted or unsubstituted alkyl group with 1-10
carbon atoms, a substituted or unsubstituted alkenyl group with 2-10
carbon atoms, a substituted or unsubstituted alkynyl group with 2-10
carbon atoms, or a substituted or unsubstituted cycloalkyl group with 3-10
carbon atoms. The unsubstituted alkyl group with 1-10 carbon atoms may,
for example, be a methyl, propyl, butyl, pentyl, hexyl, octyl, decyl
group, etc. The unsubstituted alkenyl group with 2 - 10 carbon atoms may,
for example, be a vinyl, 2-propenyl, 3-butenyl, 2-pentenyl,
2-methyl-3-pentenyl, 2-hexenyl, 5-methyl-4-hexenyl,
2,6-dimethyl-5-heptenyl group, etc. The unsubstituted alkynyl group with 2
- 10 carbon atoms may, for example, be a 2-butynyl, 3-butynyl,
1-methyl-2-pentynyl, 1-methyl-3-pentynyl, 2-hexynyl, 4-hexynyl group, etc.
The unsubstituted cycloalkyl group with 3-8 carbon atoms may, for example,
be a cyclopropyl, cyclopentyl, cyclohexyl group, etc.
Suitable substituent groups for these alkyl, alkenyl, alkynyl, and
cycloalkyl groups include halogen atoms such as fluorine and chlorine;
such lower alkoxy groups, such as methoxy, ethoxy, propoxy, and butoxy
group; a halogenoalkyl group such as trifluoromethyl; a substituted or
unsubstituted phenoxy group which has been substituted or unsubstituted
with a halogen atom or a lower alkoxy group.
The prostaglandin I.sub.2 's in formula (1) may be prepared, for example,
according to the methods described in Japan Laid-Open Patent Showa
58-150583 and Japan Laid-Open Patent Showa 57-32981.
The stable prostaglandin I.sub.2 's shown in the aforementioned formula (1)
are specifically listed as follows:
(1) Isocarbacyclin, i.e. 9(0)-methano-.DELTA..sup.6(9.alpha.) -PGI.sub.1,
(hereinafter "isocarbacyclin").
(2) 16,17,18,19,20-Pentanor-15-cyclopentylisocarbacyclin
(3) 16,17,18,19,20-Pentanor-15-cyclohexylisocarbacyclin
(4) 17,20-Dimethylisocarbacyclin
(5) 15-Deoxy-16-hydroxyisocarbacyclin
(6) 15-Deoxy-16-hydroxy-16,20-dimethylisocarbacyclin
(7) 7-Fluoroprostacyclin
(8) 7-Fluoro-16,17,18,19,20-pentanor-15cyclopentylprostacyclin
(9) 7-Fluoro-16,17,18,19,20-pentanor-15cyclohexylprostacyclin
(10) 7-Fluoro-17,20-dimethylprostacyclin
(11) 7-Fluoro-16,16-dimethylprostacyclin
(12) 7,16-Difluoroprostacyclin
(13) 15-Deoxy-16-hydroxy-7-fluoroprostacyclin
(14) 15-Deoxy-16-hydroxy-7,16-difluoroprostacyclin
(15) The methyl esters of (1) - (14)
The fat emulsions in the invention comprises, as main constituents, the
prostaglandin I.sub.2 's in formula (I), 5-50 w/v % of vegetable oil, 1-50
parts, preferably 5-30 parts, of phospholipid for 100 parts of the
vegitable oil, and an appropriate quantity of water.
The vegetable oil may be soybean oil, cotton seed oil, sesami oil, safflor
oil, and corn oil, but preferably soybean oil.
The soybean oil of choice is a refined soybean oil with high purity.
Preferably, it is the highly purified, refined soybean oil obtained by
further purifying common refined soybean oil by, for example, steam
distillation.
The phospholipid is a purified phospholipid such as egg yolk lecithin and
soybean lecithin. The phospholipid may be used which has been prepared by
fractionation using an organic solvent according to a conventional method,
that is, by slowly adding, with stirring, acetone to a crude yolk
phospholipid dissolved in a cold n-hexane-acetone mixture, collecting
insolubles by filtration, repeating the procedure of dissolution, followed
by precipitation, and finally removing the solvent by distillation. The
product comprises, as main constituents, phosphatidylcholine and
phosphatidylethanolamine. The other phospholipids may be
phosphatidylinositol, phosphatidylserine, sphingomyelin, etc.
To the fat emulsions in the invention may, where necessary, be further
added an emulsifying adjuvant, stabilizer, high molecular substance,
isotonizing agent, etc.
The emulsifying adjuvant may, for example, be up to 0.3 w/v % of fatty
acids with 6-22, preferably 12-20, carbon atoms or their pharmaceutically
acceptable salts, and so on. Either of the fatty acids with 6-22 carbon
atoms may be used if they can be added to pharmaceutical products. Such
fatty acids are either of the straight or of the branched chain; they are
preferably stearic, oleic, linolic, palmitic, linolenic, and myristic
acids. These salts may be salts with alkali metals such as sodium and
potassium, with alkaline earth metals such as calcium, and so on.
The stabilizing agent may, for example, be less than 0.5 w/v %, preferably
less than 0.1 w/v %, of the cholesterols, less than 5 w/v %, preferably
less than 1 w/v %, of phosphatidic acids, and so forth.
The high molecular substances may, for example, be 0.1-5 parts by weight,
preferably 0.5-1 part by weight, of albumin, dextran, vinyl polymers,
nonionic surface active agents, gelatin, hydroxyethyl starch, etc. for 1
part by weight of the prostaglandin I.sub.2 's.
The albumin may preferably be of human origin, and the vinyl polymers may
be polyvinylpyrrolidone, etc. The nonionic surface active agents may, for
example, be polyalkylene glycols, polyoxyalkylene copolymers, the
polyoxyalkylene derivatives of hardened castor oil, the polyoxyalkylene
derivatives of castor oil, etc.
The content of the prostaglandin I.sub.2 's in the fat emulsions may be
suitably increased or decreased according to the form of the emulsions and
applications; in general, minute quantities, for example, 1.0 mg-0.2
.mu.g/ml in the fat emulsions are sufficient.
The fat emulsions in the present invention are prepared, for example, in
the following manner: Predetermined amounts of vegetable oil,
phospholipid, the prostaglandin I.sub.2 's, and other additives are mixed
and the mixture is warmed to a solution. The solution is homogenized by,
for example, the use of a homogenizer of the high-pressure jet type, an
ultrasonic homogenizer, etc. The homogenate is further homogenized
following the addition of a necessary amount of water in order to prepare
the fat emulsions in the invention. Under preparatory conditions, the
additives such as a stabilizer and isotonizing agent may be added after
the fat emulsions have been formed.
The fat emulsions may be administered parenterally such as by injection,
most preferably intravenously. For instance, the prostaglandin I.sub.2 's
are administered intravenously by continuous infusion once a day in dose
levels of 0.01-0.1 .mu.g/kg, or 0.01-0.1 ng/kg/min. The fat emulsions in
the present invention possess very potent effects, are long-acting by
sustained release and selective for lesions; therefore, administration of
small doses enables effective treatment.
Since intravenous administration is possible, rapid onset of action can be
anticipated, drug efficacy is consistent, and since doses are small, there
is less manifestation of side effects.
Furthermore, the particles are extremely minute, and the average size of
them is less than 1.0 .mu.. The safety (stability) during storage is very
good.
Best Manner to Implement the Invention
Using the following Examples, the best methodologies to embody the present
invention are described below:
EXAMPLE 1
Preparation of a fat emulsion containing isocarbacyclin (compound I)
To 10 g of a refined soybean oil were added 1.2 g of egg yolk lecithin and
1000 .mu.g of isocarbacyclin. The mixture was heated at 60-80.degree. C.
to a solution. To this solution were added 50 ml of distilled water and
then 2.5 g of glycerol. Distilled water for injection was added to make
the solution to 100 ml. The solution was roughly emulsified in a
homomixer.
Using a Manton-Gaulin homogenizer, the crude emulsion was then further
emulsified by passing 10 times through the instrument under a first-stage
pressure of 120 kg/cm.sup.2 and a total pressure of 500 kg/cm.sup.2. There
was obtained a homogenized, finely dispersed, 10 % soybean oil-containing
fat emulsion which contained isocarbacyclin (compound I) in a final
concentration of 10 .mu.g/ml.
EXAMPLE 2
Preparation of the fat emulsions, each of which contained
16,17,18,19,20-pentanor-15-pentylisocarbacyclin (compound II),
7-fluoro-16,17,18,19, 20-pentanor-15-cyclopentylprostacyclin methyl ester
(compound III), and isocarbacyclin methyl ester (compound IV),
respectively
Using compounds II, III, and IV, the 10% soybean oil-containing fat
emulsions were prepared in the same manner as in Example 1, each of which
contained the above-mentioned compound respectively, in a final
concentration of 5 .mu.g/ml, 5 .mu.g/ml, and 10 .mu.g/ml, respectively.
EXAMPLES 5-16
Evaluation of drug activities using human platelets
The time course of the formation of cyclic AMP by the fat emulsion in the
invention was determined using human platelets so as to compare with the
time course of the compound which was not emulsified in fat. Human blood
50 ml was collected using 1 part of 3.8 % sodium citrate for 9 parts of
blood. The blood sample was centrifuged at 1300 rpm for 10 minutes. The
upper layer was taken out as PRP (platelet-rich plasma) and centrifuged at
3000 rpm for 20 minutes. The precipitate thus obtained (platelets) was
suspended in 2 ml of Tris buffered solution-saline-glucose-EDTA (TSG-EDTA)
(pH 7.4).
The fat emulsion, which had previously been transferred in a plastic tube
containing 350 .mu.l of TSG-EDIA and 50 .mu.l of platelet suspension and
incubated at 37.degree. C., was supplemented with 50 .mu.l of 5mM
isobutylmethylxanthin dissolved in saline, and 2 minutes later, reactions
were stopped with 0.5 ml of 10% trichloroacetic acid (TCA). The cells were
destroyed by thawing the lyophilized specimen in order to release
intracellular cyclic AMP. After removal of TCA with watersaturated ether,
the content of C-AMP was determined by radio immunoassay technique.
The results are summarized in Table 1. As is clear from these results, it
was demonstrated that the production capability of C-AMP by the fat
emulsion was satisfactorily maintained over time.
TABLE 1
______________________________________
Production capability and maintenance of
cyclic AMP by fat emulsions
C-AMP Relative
Time production
value
Examples Compounds (min.) (pico mol)
(%)
______________________________________
Example 5
Fat emulsion (I)
0 80 100
Example 6
" 30 100 125
Example 7
" 60 100 125
Comparison 1
Compound (I) 0 320 100
Comparison 2
" 30 200 63
Comparison 3
" 60 100 31
Example 8
Fat emulsion (II)
10 130 100
Example 9
" 30 190 146
Example 10
" 60 100 76
Comparison 4
Compound (II)
10 430 100
Comparison 5
" 30 320 74
Comparison 6
" 60 120 27
Example 11
Fat emulsion (III)
30 55 100
Example 12
" 60 30 55
Example 13
" 120 30 55
Comparison 7
Compound (III)
30 100 100
Comparison 8
" 60 90 90
Comparison 9
" 120 40 40
Example 14
Fat emulsion (IV)
30 40 100
Example 15
" 30 45 111
Example 16
" 120 28 70
______________________________________
EXAMPLE 17
Determination of platelet agglutination inhibitory action
Fifty (50) .mu.l of the fat emulsion obtained in Examples 1 and 2 was
transferred into 950 .mu.l of saline or 2 % bovine serum albumin (BSA) and
incubated at 37.degree. C. for 1, 3, and 10 minutes. Next, the ADP
platelet agglutination inhibitory action by PRP was determined using the
filtrate which had been passed through a 0.025 .mu.m filter. A known
quantity of the dilution before filtration was added to PRP and incubated
at 37.degree. C. for 1 minute. Thereafter, ADP agglutination was induced
to construct a dose-response curve. The platelet agglutination inhibitory
action was calculated from this curve. The results are presented in Table
2.
TABLE 2
______________________________________
Platelet agglutination inhibitory action
Platelet agglutination
Conditions inhibitory action (%)
Incubation
Fat Com- Fat
time elulsion pound emulsion
Compound
Dilutions
(min.) (IV) (IV) (I) (I)
______________________________________
2% BSA 1 13.7 65.4 87.5 100.1
3 23.1 NT 83.5 NT
10 27.1 56.2 90.6 107.7
Saline 10 -- -- 14.1 41.1
______________________________________
As indicated in Table 2, in the fat emulsions in the present invention, the
active ingredient of the prostaglandin I.sub.2 's is gradually released,
and thus these dosage forms provide a sustained-release delivery.
Potential applications in industry
The fat emulsions containing the prostaglandin I.sub.2 's in the present
invention possess potent effects, provide a sustained-release delivery,
and are selective for lesions. Therefore, they can provide effective
therapy in small doses and are extremely useful in the treatment of
various kinds of cardio-vascular diseases such as thrombotic diseases.
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